Limited information is available about early human placentation when many pregnancy pathologies originate. To address this knowledge gap and begin development of robust diagnostic tool to manage pregnancy complica- tions, we propose a pilot study based on a highly sensitive immune assay and our company?s base technology, Trophoblast Retrieval and Isolation from the Cervix (TRIC). TRIC uses a Pap smear, considered safe during pregnancy, to non-invasively capture hundreds of homogeneous, HLA-G-expressing, fetal cells during ongoing pregnancies at 5 to 20 weeks of gestational age (GA). We propose that these fetal cells are useful for assessing pregnancy status and assessing risk of perinatal disease in vivo. Our premise is based on data demonstrating that cells obtained by TRIC express trophoblast extravillous lineage markers (e.g., hCG, HLA-G), and have mo- lecular profiles affected by pathology. Specifically, immunocytochemical protein expression analysis of the cells demonstrates altered levels of several key proteins in pregnancies that later develop miscarriage, fetal growth restriction (FGR) or preeclampsia. We hypothesize that, based on robust data obtained in our published studies, that AFP and PGF levels are significantly altered in EVT cells from pregnancies linked to FGR, a key indicator of placenta-based perinatal disorders. Therefore, we will rigorously determine if expression of two proteins iden- tified in our preliminary studies in fetal cells obtained by TRIC correlate with fetal growth rates, considering fetal sex and other relevant factors. The specific aim of this application is to establish the relationship of cervical extravillous trophoblast biomarker levels to pregnancy outcomes. We will quantify AFP and PGF in EVT cells obtained by TRIC in the first trimester, comparing cohorts of subjects with normal term pregnancies to those with adverse outcomes associated with low birth weight for GA at birth. This goal will be accomplished in four milestones. Milestone 1, we will develop a high-sensitivity immune assay to quantify AFP and PGF levels in trophoblast cells. Milestone 2, we will build a biobank of 100 specimens of trophoblast cells isolated by TRIC from pregnant patients in the first trimester, along with de-identified medical records to determine pregnancy outcomes. Milestone 3, the immune assay will be optimized with patient trophoblast samples. Milestone 4, the remaining patient trophoblast samples will be assayed for AFP and PGF levels and compared to patient out- comes, particularly reduced birth weight for gestational age. These experiments will validate the testing platform and provide preliminary data for a larger clinical study to establish a perinatal test for maternal-fetal disorders. Our long-term goal is to develop and commercialize products for early diagnosis of perinatal pathologies using EVT cells obtained safely during ongoing pregnancies through TRIC.